PI3k / Akt 通路在大戟大枣汤抗不同分子表型乳腺癌中的调控作用
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1.齐齐哈尔医学院 医药科学研究院,黑龙江 齐齐哈尔 161006;2.齐齐哈尔医学院 药学院,黑龙江 齐齐哈尔 161006;3.齐齐哈尔医学院 公共卫生学院,黑龙江 齐齐哈尔 161006;4.齐齐哈尔医学院附属第五医院,黑龙江 大庆 163001;5.齐齐哈尔医学院附属第三医院,黑龙江 齐齐哈尔 161099

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R-33

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Effects of decoction of Euphorbia fischeriana Steud. and jujuba against breast cancer of different molecular phenotypes via PI3k / Akt pathway
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1. Research Institute of Medical Science and Pharmacy, Qiqihaer Medical University, Qiqihaer 161006, China.2. School of Pharmacy, Qiqihaer Medical University, Qiqihar 161006. 3. School of Public Health, Qiqihaer Medical University,Qiqihaer 161006. 4. the Fifth Affiliated Hospital of Qiqihaer Medical University, Daqing 163001.5. the Third Affiliated Hospital of Qiqihaer Medical University, Qiqihar 161099

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    摘要:

    目的 探讨 PI3k / Akt 通路在大戟大枣汤( decoction of Euphorbia fischeriana Steud. and jujuba,DEFSJ)抗雌激素受体(ER)阴性(-)和 ER 阳性(+)乳腺癌中的调控作用,为乳腺癌的针对性治疗提供参考。 方法制备 DEFSJ 提取液,采用 UHPLC-Triple Quad 仪进行质控分析。 DEFSJ 含药血清(containing serum,CS)依照体外血清药理学方法进行获取,乳腺癌(ER-)MDA-MB-453 和(ER+)MCF-7 细胞分别用不同浓度 DEFSJ-CS 干预 48 h。流式细胞术检测细胞周期分布,DNA ladder 实验评估细胞凋亡程度,Western blot 法检测 PI3k / Akt 通路相关蛋白表达,实时定量聚合酶链反应(qRT-PCR)法检测 FoxO3a、FoxO1a 及 Bim mRNA 表达;激光共聚焦显微镜观察 FoxO3a蛋白核转位。 结果 UPLC 对 5 批 DEFSJ 提取物进行了分析,结果表明制备工艺可行,质量可控,保证了药理学实验结果的准确性。 体外实验中发现,DEFSJ-CS 能阻断细胞在 G2 / M 期(P<0. 05,P<0. 01);DNA ladder 实验中呈现典型的细胞凋亡梯带;与阴性对照组比较,DEFSJ-CS 能使 p-PI3k、p-Akt、p-FoxO3a 及 p-FoxO1a 蛋白表达降低(P<0. 05,P<0. 01),Bim 蛋白表达升高(P<0. 05);使 FoxO3a、FoxO1a mRNA 表达降低(P<0. 05,P<0. 01),Bim mRNA 表达升高(P<0. 05,P<0. 01);并能使细胞中 FoxO3a 蛋白呈现显著的核转位变化;且各数据结果均显示 DEFSJ-CS 对(ER-)MDA-MB-453 细胞比对(ER+)MCF-7 细胞作用效果更好。 结论 PI3k / Akt 通路参与了大戟大枣汤抗乳腺癌作用的调控,作用效果因乳腺癌表型差异而不同。

    Abstract:

    Objective To explore the effect of decoction of Euphorbia fischeriana Steud. and jujuba (DEFSJ) against estrogen receptor (ER) negative (-) and ER positive (+) breast cancer via the PI3k / Akt pathway, and to provide a reference for the targeted treatment of breast cancer. Methods DEFSJ extract was prepared and analyzed using UHPLCTriple Quad. DEFSJ containing serum (CS) was prepared via a serum pharmacology method . Different concentrations of DEFSJ-CS were applied to (ER-) MDA-MB-453 and (ER+) MCF-7 breast cancer cells in vitro for 48 h. The distribution of cells in different stages of the cellular cycle was evaluated using a Flow cytometer. DNA ladder assay was used to assess the degree of apoptosis, and the expression of PI3k / Akt pathway-related proteins was evaluated by Western blot assay. The expression of FoxO3a, FoxO1a, and Bim mRNA was detected by Real-time quantitative PCR. Nuclear transposition of FoxO3a protein was analysed using a confocal laser microscopy. Results Five batches of DEFSJ extract were analyzed using UPLC, and the result showed that the preparation technology was feasible and the quality was controllable, ensuring the accuracy of the pharmacological experiment result. DEFSJ-CS blocked cells in the G2 / M phase (P<0. 05, P<0. 01).Cells treated with DEFSJ-CS displayed the typical apoptotic ladder in the DNA ladder experiment. Compared with the negative control cells, the DEFSJ-CS group cells had decreased protein expression of p-PI3k, p-Akt, p-FoxO3a, and pFoxO1a (P<0. 05, P<0. 01); increased protein expression of Bim (P<0. 05); decreased mRNA expression of FoxO3a and FoxO1a ( P<0. 05,P<0. 01); increased mRNA expression of Bim ( P<0. 05,P<0. 01); and enhanced nuclear translocation of FoxO3a protein. The data showed that DEFSJ-CS had a stronger effect on (ER-) MDA-MB-453 cells than (ER+) MCF-7 cells. Conclusions The regulatory effect of DEFSJ extract on anti-breast cancer involves the PI3k / Akt pathway, and the effect varies with phenotypic differences.

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马立威,姚宏玉,陈 哲,倪世宇,陈 颂,李 京,刘吉成. PI3k / Akt 通路在大戟大枣汤抗不同分子表型乳腺癌中的调控作用[J].中国比较医学杂志,2024,34(3):27~35.

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  • 收稿日期:2023-04-19
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  • 在线发布日期: 2024-05-29
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